The development and validation of a rapid method of RRLC has been carried out to determine the phenolic composition of winemaking by-products (pomaces, seeds, skins and stems). Thirty-one phenolic compounds belonging to three groups (flavanols, flavonols and phenolic acids) have been identified by use of standards and mass spectrometric detection, and quantified by using the corresponding external standard calibration plot, in a 16-min run. The validation was realized calculating the repeatability, the reproducibility and the limits of detection (LOD) and quantification (LOQ), from standards solutions. The limits of detection and quantification were in the range of 0.16-1.09 and 0.52-3.63mg/L, respectively, and good repeatability (R.S.D. values <1.5%) and reproducibility (R.S.D. values <5.5%) were found. Results confirmed that the method is effective and suitable for determination of phenolic compounds in winemaking by-products. Seeds, skins, stems and pomaces exhibited a different qualitative and quantitative phenolic profile and different antioxidant activities.